To further investigate whether the neuroprotective effects of intracerebroventricular injection of calycosin against CIRI in rats are associated with the HMGB1/NLRP3-mediated pyroptosis signaling pathway, we extracted total RNA and protein from the ischemic penumbra of brain tissues from each group of rats at 24 h reperfusion following 1.5 h of cerebral ischemia and conducted qRT-PCR and Western blot analyses. This evidence concerns the gene HMGB1 and brain ischemia.